生物毒素的脱毒技术及药物研究进展

作为一种特殊的生物源化学物质,生物毒素对农产品、人畜、环境等有不可忽视的健康隐患,但由于其结构与功能作用的多样性,其对于生物化学、医药学、环境生态学等又具有重要意义。本研究就生物毒素的来源分类、危害、脱毒技术及生物毒素的应用进行综述,以期为进一步生物毒素的监管与开发利用提供理论参考。     

Modelling and fabrication of optimum structure of novel interdigital sensors for food inspection

Modelling of an interdigital sensor‐based sensing system for the detection of dangerous contaminated marine biotoxin in seafood has been carried out. Three interdigital sensors having new electrode configurations have been analysed using COMSOL Multiphysics. All sensors were designed to have an equal number of electrodes with the same sensing area. Based on the maximum sensitivity, the best electrode configuration has been chosen. Experiments on raw seafood contaminated with domoic acid have been conducted using the new fabricated sensor and very good results were obtained. Copyright © 2011 John Wiley & Sons, Ltd.     

食品中致癌性生物毒素检测标准概述

生物毒素具有较高的生物毒性,摄入受到生物毒素污染的食品,会对大众健康会造成极大危害。由于该类物质的高风险性,许多国际组织或国家制定了诸多有关食品中致癌性生物毒素的法规,对其进行严格控制。本文梳理了主要国际组织和国家关于致癌性生物毒素限量和检测方法,并与我国现行标准进行了比较,对发现的问题提出了一些建议,希望为该领域相关检测机构或企业进行致癌性生物毒素检测时选择方法提供参考。     

固相萃取-超高效液相色谱-串联质谱法快速测定农产食品中14种真菌类生物毒素

目的建立超高效液相色谱-串联质谱法同时测定农产食品中14种真菌类生物毒素的分析方法。方法样品经乙腈-水溶液提取后,Mycosep~226多功能净化柱净化后,利用超高效液相色谱-串联质谱测定,正负离子同时扫描,多反应监测模式同时测定14种生物毒素,外标法定量。结果该方法在0.3~20μg/L的浓度范围内的线性相关系数较好(r~20.9990),定量限范围为0.3~1.5μg/kg,3个水平的平均加标回收率范围为71.7%~96.1%,相对标准偏差范围为3.35%~7.93%。结论该方法操作简单、快速准确、干扰小,适用于农产食品中14种生物毒素的检测。     

Determination of the toxic variability of lipophilic biotoxins in marine bivalve and gastropod tissues treated with an industrial canning process

Contamination of shellfish with lipophilic marine biotoxins (LMB), pectenotoxins (PTXs), yessotoxins (YTXs) and okadaic acid (OA) toxin groups in southern Chile is a constant challenge for the development of miticulture considering the high incidence of toxic episodes that tend to occur. This research is focused on using methodologies for assessing the decrease in toxins of natural resources in Chile with high value, without altering the organoleptic properties of the shellfish. The species were processed through steaming (1 min at 121°C) and subsequent canning (5 min at 121°C). Changes in the profiles of toxins and total toxicity levels of LMB in endemic bivalves and gastropods were determined using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The total reduction of toxicity (≈ 15%) was not related to the destruction of the toxin, but rather to the loss of LMB on removing the shells and packing media of canned products (***p < 0.001). Industrial processing of shellfish reduces LMB contents by up to 15% of the total initial contents, concomitant only with the interconversion of PTX-group toxins into PTX-2sa. In soft bottom-dwelling species with toxicities beyond the standard for safe human consumption (≥ 160 μg OA-eq kg^–1), toxicity can be reduced to safe levels through industrial preparation procedures.     

Screening for the presence of lipophilic marine biotoxins in shellfish samples using the neuro-2a bioassay

The neuro-2a bioassay is considered as one of the most promising cell-based in vitro bioassays for the broad screening of seafood products for the presence of marine biotoxins. The neuro-2a assay has been shown to detect a wide array of toxins like paralytic shellfish poisons (PSPs), ciguatoxins, and also lipophilic marine biotoxins (LMBs). However, the neuro-2a assay is rarely used for routine testing of samples due to matrix effects that, for example, lead to false positives when testing for LMBs. As a result there are only limited data on validation and evaluation of its performance on real samples. In the present study, the standard extraction procedure for LMBs was adjusted by introducing an additional clean-up step with n-hexane. Recovery losses due to this extra step were less than 10%. This wash step was a crucial addition in order to eliminate false-positive outcomes due to matrix effects. Next, the applicability of this assay was assessed by testing a broad range of shellfish samples contaminated with various LMBs, including diarrhetic shellfish toxins/poisons (DSPs). For comparison, the samples were also analysed by LC-MS/MS. Standards of all regulated LMBs were tested, including analogues of some of these toxins. The neuro-2a cells showed good sensitivity towards all compounds. Extracts of 87 samples, both blank and contaminated with various toxins, were tested. The neuro-2a outcomes were in line with those of LC-MS/MS analysis and support the applicability of this assay for the screening of samples for LMBs. However, for use in a daily routine setting, the test might be further improved and we discuss several recommended modifications which should be considered before a full validation is carried out.     

Recent advancements in LC-MS based analysis of biotoxins: Present and future challenges

There has been a rising concern regarding the harmful impact of biotoxins, source of origin, and the determination of the specific type of toxin. With numerous reports on their extensive spread, biotoxins pose a critical challenge to figure out their parent groups, metabolites, and concentration. In that aspect, liquid chromatography-mass spectrometry (LC-MS) based analysis paves the way for its accurate identification and quantification. The biotoxins are ideally categorized as phytotoxins, mycotoxins, shellfish-toxins, ciguatoxins, cyanotoxins, and bacterial toxins such as tetrodotoxins. Considering the diverse nature of biotoxins, both low-resolution mass spectrometry (LRMS) and high-resolution mass spectrometry (HRMS) methods have been implemented for their detection. The sample preparation strategy for complex matrix usually includes “QuEChERS” extraction or solid-phase extraction coupled with homogenization and centrifugation. For targeted analysis of biotoxins, the LRMS consisting of a tandem mass spectrometer operating in multiple reaction monitoring mode has been widely implemented. With the help of the reference standard, most of the toxins were accurately quantified. At the same time, the suspect screening and nontarget screening approach are facilitated by the HRMS platforms during the absence of reference standards. Significant progress has also been made in sampling device employment, utilizing novel sample preparation strategies, synthesizing toxin standards, employing hybrid MS platforms, and the associated data interpretation. This critical review attempts to elucidate the progress in LC-MS based analysis in the determination of biotoxins while pointing out major challenges and suggestions for future development.     

超高效液相色谱-四级杆/静电场轨道阱高分辨质谱联用快速测定水产品及干制水产品制品中的116种农药和24种生物毒素残留

采用了一种新型脂肪吸附剂去除样品基质中脂肪和磷脂等杂质的干扰,利用超高效液相色谱-四级杆/静电场轨道阱质谱法(UPLC-Q Exactive Orbitrap MS)的同时定性定量功能,建立了水产品及干制水产品制品中116种农药和24种生物毒素残留量的检测方法。样品前处理采用QuEChERS方法,经乙腈/水(90:10,V/V)溶液提取,高效基质脂肪吸附剂(EMR-Lipid)净化,平行定量浓缩仪浓缩,C18色谱柱分离,5 mmol甲酸水溶液(含0.1%甲酸铵)和5 mmol甲酸甲醇溶液(含0.1%甲酸铵)梯度洗脱;质谱数据采集使用Q Exactive高分辨质谱的Full MS/dd-MS2监测模式,以Full MS一级质谱全扫描提取母离子精确质量数所得的色谱峰面积进行定量,以保留时间和dd-MS2数据依赖子离子扫描所得的二级子离子质谱图进行定性确证。140种目标物的精确质量数偏差不大于3×10^-6,浓度与母离子峰面积的线性关系良好,相关系数≥0.991,检出限为0.02~0.4μg/kg。基质加标回收率在70.1%~109.1%之间,相对标准偏差(RSD)为1.0%~14.1%。该方法学结果满足GB/T 27417-2017《合格评定化学分析方法确认和验证指南》的技术要求。本方法具有操作简单快捷、灵敏度高等优点。     

超高效液相色谱-串联质谱法在食品安全检测中的应用

食品安全与人们的生活和健康密切相关, 关系到社会和谐稳定和经济平稳运行。高效的检测技术方法, 对保障食品安全具有重要意义。超高效液相色谱-串联质谱(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS), 以其高效、快速、灵敏、高选择性和高通量等优点, 为食品安全检测提供了一个重要的工具。该技术与QuEChERS、液-液微萃取、分散固相萃取、免疫亲和柱净化、碳纳米管纯化及柱前衍生化等多种样品制备技术的结合, 使其在食品样品复杂基质中痕量成分检测中发挥出独特的技术优势。本文综述了UPLC-MS/MS在食品农药兽药残留检测、食品生物毒素检测、食品加工危害物检测和食品过敏原检测等食品安全检测领域的应用和进展, 并对其未来应用潜力和发展前景进行了展望, 以期为UPLC-MS/MS技术方法在食品安全检测领域更好地推广应用提供参考。